Using the Landsat TM imagery and land use data of Shenzhen in Oct. 2009, this paper studied the relationship between land use structure and spatial differentiation of thermal environment, by land surface temperature (LST) retrieval, temperature zoning and overlay mapping. Besides, investigation of plots in three temperature partitions and analysis of landscape metrics were also used to study the relationship between landscape characteristics and LST. The results showed that, 1) there were significant variation in both land use structure and spatial pattern of different temperature partition. The landscape of mid-temperature partition was highly fragmented and patch shape was most complex. In cooling-island partition, heterogeneity of landscape was low and patch shape were relatively simple with an aggregated structure. 2) relationships between spatial pattern and LST also differed among the three partitions. The landscape-level configuration metrics were all significantly negatively correlated to LST in heat-island partition , however, they displayed a positive relationship in cool-island partition. 3) In greenspace and water area, several patch-level landscape metrics displayed an inconsistent relationship among three partitions, suggested there existed a threshold character in cooling effect of ecological land uses. Research on relationship between urban landscape pattern and LST can help us understand the influence of the land use pattern on surface thermal environment, and it also provide support for studies about optimization of landscape pattern to ease heat island effect.